Characterizing eye-gaze positions of people with severe motor dysfunction: Novel scoring metrics using eye-tracking and video analysis.

Nonverbal communication with people who have physical disabilities is difficult. Eye-tracking technologies have recently been developed and applied to help people with physical disabilities in their communication. However, the eye-gaze patterns of people with severe motor dysfunction (SMD) have not been analyzed in detail. To clarify characterization of people with SMD, we aimed to develop gaze position-based evaluation metrics and analyze detailed eye-gaze patterns of people with SMD. We developed two new scoring metrics: (1) saliency score based on three saliency maps-spectral residual (SR), fine grained (FG), and motion (Mo); and (2) the distance score, which represents to what extent people can chase an object in a video. The evaluation was performed on 102 participants, consisting of 35 subjects with profound intellectual and multiple disabilities (PIMD; SMD with IQ < 20), 19 with severe physical disabilities (SPD; SMD with IQ ≥ 20), and 48 healthy individuals. We observed that two saliency scores (SR and FG) and the distance score showed significant differences between the PIMD/SPD and healthy groups for the entire video, whereas Mo scores did not. Moreover, the distance score was analyzed separately for each scene, where scenes were categorized into three patterns-running, explanation, and hiding-according to the behavior of the moving objects. In the SPD and healthy groups, the explanation scenes accounted for the highest percentage of all scenes with the best distance score (63.6% and 61.9%, respectively), whereas in the PIMD group, the running scenes accounted for the highest percentage (54.5%). In conclusion, the new metrics were successful in quantitatively assessing the gaze responsiveness of people with SMD, which could not be assessed using a conventional metric, gaze-acquisition time. This study is expected to expand the possibilities of nonverbal communication using eye-tracking devices for people with SMD.

Obstetric complication-associated ANXA5 promoter polymorphisms may affect gene expression via DNA secondary structures.

Recent findings have highlighted the possibility that polymorphisms within the annexin A5 gene (ANXA5) promoter contribute to the etiology of various obstetric complications. However, the underlying mechanisms are unknown. The M2 haplotype of the ANXA5 shows lower activity and less expression of ANXA5 mRNA. This gene promoter region has a motif that potentially forms a G-quadruplex structure. In vitro G-quadruplex propensity estimated by circular dichroism indicated that the M2 haplotype oligonucleotide manifested a decreased potential for G-quadruplex formation. In addition, in vivo G-quadruplex formation of the promoter region was evidenced by the presence of single-stranded DNA shown by sodium bisulfite treatment of placental genomic DNA. Comparative analysis indicated less potential in the M2 allele than the major allele. Promoter activity of the two haplotypes determined by luciferase reporter analysis correlated with the estimated G-quadruplex propensity. Our data lend support to the developing paradigm that genomic variation affects gene expression levels via DNA secondary structures leading to the disease susceptibility.

The experience of parents of adult sons with Duchenne muscular dystrophy regarding their prolonged roles as primary caregivers: a serial qualitative study.

PURPOSE: Mechanical ventilation has allowed a greater number of patients with Duchenne muscular dystrophy (DMD) to transition into adulthood. However, the role of a child's parent as a caregiver lasts throughout the child's lifetime. We explored parents' experiences of prolonged caregiving using serial interviews, analyzed using constructivist grounded theory. MATERIALS AND METHODS: Fourteen parents (average age 53.9 years) with sons with DMD (average age 23.2 years) were interviewed two to four times, over a 3-year period. Data were analyzed using a grounded theory approach. RESULTS: Two categories of responses were defined as strengths, and four as weaknesses. The strengths were related to family member support and confidence in parenting ability. The weaknesses were related to the anticipation of aging with the ongoing burden of caring for adult sons, regrets, sharing of responsibility versus having a fixed role as the primary caregiver, and economic burden. The weaknesses became more pronounced as the duration of caregiving increased. Parents' acceptance of and immobilization in their role of primary caregiver led to prolonged derivative dependency. CONCLUSION: Practical support for parental caregivers, who experience a marked increase in the duration of their caregiving role while facing their own aging-related challenges, are required. Implications for Rehabilitation Children with DMD are living longer and are transitioning into adulthood; a successful transition involves becoming as independent as possible and maintaining a positive sense of personal identity. Despite entering adulthood, the parental caregiver's caregiving role continues. Rehabilitation professionals, who are able to provide long-term, continued support from childhood into adulthood, should be aware that parental caregivers' weakness are exacerbated as the duration of caregiving increases. Families affected by DMD require multifaceted support that should include support for the parental caregiver.

Caregivers' lived experience in trying to read slight movements in a child with severe brain injury: A phenomenological study.

AIMS AND OBJECTIVES: To explore caregivers' lived experience of reading slight movements of a child with severe brain injury. BACKGROUND: Despite increased need, the development of individual care for children with severe brain injuries has been prevented by their severe physical state and the poor reproducibility of their movements. In addition to a lack of evidence on the motor characteristics of patients with severe brain injury with multiple disabilities, their own development contributes to increasing variability in their states. Thus, caregivers are compelled to rely on their experiences, which have not been academically explored. DESIGN: A qualitative study based on van Manen's method of hermeneutic phenomenology. METHODS: Data were obtained through twenty-one 3-hr observation sessions and five 15- to 45-min group interviews. We observed a child (called AK) with severe brain injury and his 61 caregivers, and conducted group interviews with 28 caregivers. We focused on caregivers' experiences of reading AK's slight movements. The data were interpreted based on van Manen's hermeneutic phenomenological approach. RESULTS: Four themes emerged as caregivers' experience in trying to read AK's slight movements. By considering "AK's physical state and his slight movements" and discovering "caregivers' 'sense of uncertainty' about AK's slight movements," caregivers could decipher "AK's multiple slight movements." "Sharing" was found as a necessary aspect of these other three themes of reading AK's slight movements. CONCLUSIONS: We presented caregivers' experiences as related to these four themes in their efforts to read the slight movements of AK. Due to AK's slight movements with poor reproducibility, "sharing" was necessary to read AK's slight movements, as it exposes caregivers' lived experience to the interpretation of multiple caregivers. RELEVANCE TO CLINICAL PRACTICE: These four themes may be useful for assessing, guiding and promoting caregivers' use of sharing when reading the slight movements of children with severe brain injury.

Longitudinal burnout-collaboration patterns in Japanese medical care workers at special needs schools: a latent class growth analysis.

The present study aimed to identify and characterize potential burnout types and the relationship between burnout and collaboration over time. Latent class growth analysis and the growth mixture model were used to identify and characterize heterogeneous patterns of longitudinal stability and change in burnout, and the relationship between burnout and collaboration. We collected longitudinal data at three time points based on Japanese academic terms. The 396 study participants included academic teachers, yogo teachers, and registered nurses in Japanese special needs schools. The best model included four types of both burnout and collaboration in latent class growth analysis with intercept, slope, and quadratic terms. The four types of burnout were as follows: low stable, moderate unstable, high unstable, and high decreasing. They were identified as involving inverse collaboration function. The results indicated that there could be dynamic burnout types, namely moderate unstable, high unstable, and high decreasing, when focusing on growth trajectories in latent class analyses. The finding that collaboration was dynamic for dynamic burnout types and stable for stable burnout types is of great interest. This was probably related to the inverse relationship between the two constructs.

Role development of nurses for technology-dependent children attending mainstream schools in Japan.

PURPOSE: To describe the role development of nurses caring for medical technology-dependent children attending Japanese mainstream schools. DESIGN AND METHODS: Semi-structured interviews with 21 nurses caring for technology-dependent children were conducted and analyzed using the modified grounded theory approach. RESULTS: Nurses developed roles centered on maintaining technology-dependent children's physical health to support children's learning with each other, through building relationships, learning how to interact with children, understanding the children and the school community, and realizing the meaning of supporting technology-dependent children. PRACTICE IMPLICATIONS: These findings support nurses to build relationships of mutual trust with teachers and children, and learn on the job in mainstream schools.

Becoming a back-up carer: parenting sons with Duchenne muscular dystrophy transitioning into adulthood.

The population of adults with Duchenne muscular dystrophy is increasing rapidly. However, information for individuals with DMD and their parents about the transition to adulthood is lacking; young adult sons and their parents may struggle to maintain smooth family functioning and well-being during this period. This study examined the process of change in parental behaviors during their son's transition. The participants were 18 parents with sons aged 15-30 years. Data were obtained from semi-structured interviews and analyzed using a grounded theory approach. Eleven categories of behaviors were identified across three domains: emotional, physical, and determination. The changes made by parents were directed toward becoming a back-up carer: letting go of some control but still being active participants in their sons' lives. We identified several issues important for well-being in the transition period: psychological support, the aging of the parents (the primary caregivers) and the concomitant emergency and specialized care needs, and parents' intervention in the self-determination of adult sons with DMD. The findings of this study may provide a rationale to advocate for policies to improve support for parents of sons with DMD transitioning to adulthood and provide information to help parents in their role as primary care providers.

Independent living with Duchenne muscular dystrophy and home mechanical ventilation in areas of Japan with insufficient national welfare services.

In Japan, there is no national 24-hour home care system for people with severe impairments. Despite this fact, a small number of people with Duchenne muscular dystrophy on home mechanical ventilation pursue independent living. Therefore, our aim was to better understand the process by which these individuals arrived at this goal for independence (i.e., choosing to live at home in Japan instead of in special sanatoriums that provide sufficient support and care). Twenty-one participants were interviewed in 2011 and 2013. The interviews were recorded, transcribed, and analysed following a grounded theory approach. These individuals placed particular emphasis on their personal choice regarding where and how they live as well as on whom they depend. Therefore, the core element underlying participants' goals for independent living was self-reliant independency. To improve their social inclusion, the strategies used by the participants to retain their autonomy in an underdeveloped Japanese welfare system by establishing relationships with people in their communities can prevent them from experiencing social isolation. This could serve as an example to their counterparts in other countries.

Decreased expression of apelin in placentas from severe pre-eclampsia patients.

OBJECTIVE: It is well documented that anti-angiogenic factors are likely to play essential roles in the etiology of pre-eclampsia. Apelin is a small peptide that may potentially act as an angiogenic factor. The expression of apelin was examined at the RNA and protein levels in this study. METHODS: We compared the expression of apelin, examined using quantitative reverse-transcription polymerase chain reaction, western blotting, enzyme-linked immunosorbent assay and immunostaining, between pre-eclamptic patients and normotensive controls. RESULTS: Apelin messenger RNA is significantly decreased in pre-eclamptic placentas compared with normotensive pregnancies (p<0.05). Apelin protein levels are also lower in pre-eclamptic placentas than the controls but higher in the maternal circulation in pre-eclampsia patients. Immunohistochemical signals for apelin and its receptor APJ were detected mainly in the cytoplasm of syncytiotrophoblasts in chorionic villi and trophoblast-lineage cells in the decidua of term placentas. In early gestation, stronger APJ signals were observed at the cellular membrane. CONCLUSIONS: A functional role of the apelin--APJ system is likely in early gestation, and this raises the possibility that a dysfunctional apelin--APJ system contributes to the onset of pre-eclampsia via decreased angiogenic activity in placental implantation.

Comparative gene expression profiling of placentas from patients with severe pre-eclampsia and unexplained fetal growth restriction.

BACKGROUND: It has been well documented that pre-eclampsia and unexplained fetal growth restriction (FGR) have a common etiological background, but little is known about their linkage at the molecular level. The aim of this study was to further investigate the mechanisms underlying pre-eclampsia and unexplained FGR. METHODS: We analyzed differentially expressed genes in placental tissue from severe pre-eclamptic pregnancies (n = 8) and normotensive pregnancies with or (n = 8) without FGR (n = 8) using a microarray method. RESULTS: A subset of the FGR samples showed a high correlation coefficient overall in the microarray data from the pre-eclampsia samples. Many genes that are known to be up-regulated in pre-eclampsia are also up-regulated in FGR, including the anti-angiogenic factors, FLT1 and ENG, believed to be associated with the onset of maternal symptoms of pre-eclampsia. A total of 62 genes were found to be differentially expressed in both disorders. However, gene set enrichment analysis for these differentially expressed genes further revealed higher expression of TP53-downstream genes in pre-eclampsia compared with FGR. TP53-downstream apoptosis-related genes, such as BCL6 and BAX, were found to be significantly more up-regulated in pre-eclampsia than in FGR, although the caspases are expressed at equivalent levels. CONCLUSIONS: Our current data indicate a common pathophysiology for FGR and pre-eclampsia, leading to an up-regulation of placental anti-angiogenic factors. However, our findings also suggest that it may possibly be the excretion of these factors into the maternal circulation through the TP53-mediated early-stage apoptosis of trophoblasts that leads to the maternal symptoms of pre-eclampsia.

Impact of indoleamine 2,3-dioxygenase on the antioxidant system in the placentas of severely pre-eclamptic patients.

Indoleamine 2,3-dioxygenase (IDO) is the rate limiting enzyme of the kynurenine pathway that degrades L-tryptophan, but a wider range of functions have now been proposed for this enzyme, including antioxidant activity. Our previous study revealed that reduced IDO expression in the placenta induces defective feto-maternal immuno-tolerance leading to the onset of pre-eclampsia. In our present study, we assessed the effects of low placental IDO activity as an antioxidant. The placental levels of 8-hydroxy-2'-deoxy-guanosine (8-OHdG), a maker for oxidative damage to DNA, were significantly higher in pre-eclamptic than normotensive pregnancies (P<0.05). Immunohistochemical signals of 8-OHdG were detected mainly in syncytiotrophoblasts and vascular endothelial cells, and co-localized with those for IDO. Furthermore, a significant inverse correlation was found between the IDO activity and 8-OhdG levels. These results show that oxidative stress is associated with decreased IDO activity in the pre-eclamptic placenta and suggest an impact of low IDO activity other than immune modulation in promoting the onset of this disorder.

Polymorphisms in the annexin A5 gene promoter in Japanese women with recurrent pregnancy loss.

Recent findings have raised the possibility that polymorphisms within the annexin A5 gene (ANXA5) promoter contribute to the etiology of recurrent pregnancy loss (RPL). In our present study, 243 Japanese women who had suffered more than three fetal losses and a group of 119 fertile controls were genotyped for four ANXA5 gene promoter single-nucleotide polymorphisms (SNPs; SNP1-4: g.-467G >A, g.-448A>C, g.-422T>C, g.-373G>A) previously reported to be associated with this disorder. An additional two SNPs located within the 5'-untranslated region of the ANXA5 (SNP5 and 6: g.-302T>G, g.-1C>T) were also evaluated. Our case--control study revealed that the minor allele was significantly more frequent in the RPL group than controls for all six of these SNPs, among which SNP5 showed the highest significance (P= 0.002). As with the M2 haplotype for SNP1-4 (A-C-C-A) for a western population in previous reports, a haplotype comprising all of the minor alleles for SNP1-6 (A-C-C-A-G-T), the third major haplotype in the Japanese population, showed a significantly higher frequency in our current RPL subjects than in controls (P= 0.025). In addition, the second major haplotype (G-A-T-G-G-C) was found to confer a significant risk of RPL (P= 0.036), implicating SNP5 as a major risk determinant for this disease. Our present findings support the hypothesis that genomic variations within the ANXA5 gene upstream region impact upon the disease susceptibility to RPL. Our data indicate that SNP5 is a novel risk factor for this disease in the Japanese population.

Genetic variation in the indoleamine 2,3-dioxygenase gene in pre-eclampsia.

PROBLEM: To investigate the contribution of genomic variations in the indoleamine 2,3-dioxygenase (IDO) gene to the onset of pre-eclampsia. METHOD OF STUDY: We examined sequence variations in the IDO1 gene using placental genomic DNA from 35 pre-eclamptic patients and 32 normotensive pregnant women. RESULTS: A case-control study revealed that none of the common variants influences the risk of disease. Sequencing of each IDO1 exon in diseased subjects revealed rare variants. This variation, c.-147_150delGAAA, was located within the 5'-untranslated region of the IDO1 gene, and its homozygote was identified only in pre-eclamptic subjects. However, despite the low levels of IDO expression and enzyme activity in the c.-147_150delGAAA homozygote, reporter assays indicated that this variation does not affect gene expression. CONCLUSION: Our findings indicate that genetic alteration of fetal IDO gene does not appear to be a primary cause of pre-eclampsia.

CD9 gene variations are not associated with female infertility in humans.

BACKGROUND/AIMS: To determine whether genetic alterations in the CD9 gene are associated with female infertility in humans. METHODS: We sequenced the entire coding region of this gene in 86 Japanese women with unexplained infertility and further conducted a case-control study of six tagging single nucleotide polymorphisms (SNPs) in this gene using an additional 164 samples obtained from a fertile control group. RESULTS: No disease-causing mutation in the CD9 gene was evident in these samples and no significant association between the tagging SNPs and the studied cohort was identified. CONCLUSIONS: Our findings do not support the hypothesis that genetic alterations of the CD9 gene cause female infertility in humans.

Analysis of nitric oxide metabolism as a placental or maternal factor underlying the etiology of pre-eclampsia.

BACKGROUND: Defective nitric oxide (NO)-mediated vasodilation is widely regarded as an underlying cause of hypertension in pre-eclampsia, although there are also arguments against this hypothesis. METHODS: We examined both the mRNA levels and the presence of a Glu298Asp substitution in the NO synthase (NOS) gene, as well as the NO metabolite concentration, in placentas and maternal sera from women with pre-eclampsia and in normotensive pregnant controls (25-40 vs. 24-41 weeks of gestation). RESULTS: Pre-eclamptic and control placentas did not show any significant differences in their NO metabolite levels or their NOS expression levels as measured by quantitative RT-PCR. In addition, we did not find any association between pre-eclampsia and the occurrence of the Glu298Asp amino acid substitution in the NOS gene. In contrast, high maternal circulating NO metabolites were evident in severe pre-eclampsia (p < 0.0001). Although a positive correlation between circulating NO metabolites and blood pressure was not observed, uterine artery resistance measured by ultrasound was found to positively correlate with the maternal NO levels. CONCLUSIONS: Our current data suggest that an altered placental NOS pathway is unlikely to be the primary cause of pre-eclampsia and that the activation of this pathway is possibly in response to maternal symptoms.

Increased levels of pregnancy-associated plasma protein-A2 in the serum of pre-eclamptic patients.

Pregnancy-associated plasma protein-A and -A2 (PAPP-A and -A2) are proteases that cleave insulin-like growth factor-binding proteins (IGFBPs), resulting in local activation of IGF signaling pathways. Here, we examined PAPP-A and -A2 mRNA and protein levels in placenta and maternal sera from women with pre-eclampsia and compared them with samples from uncomplicated pregnancy. PAPP-A2 but not PAPP-A mRNA and protein were elevated in pre-eclamptic placenta (P < 0.01). PAPP-A2 is normally produced in placental syncytiotrophoblast cells and maternal decidua. PAPP-A2 in syncytiotrophoblast cells was dramatically increased in pre-eclampsia. Maternal serum concentrations of PAPP-A2 but not PAPP-A were also significantly elevated in pre-eclampsia as compared with uncomplicated pregnancy. mRNA levels of IGFBP5, a specific substrate for PAPP-A2 protease activity, were also significantly increased, suggesting a potential role for IGFBP5 in fetal and placental growth suppression during pre-eclampsia. However, IGFBP5 protein levels were not increased in placenta from pre-eclampsia, possibly due to cleavage by up-regulated PAPP-A2. These data might imply that PAPP-A2 may be up-regulated in pre-eclamptic pregnancy to compensate for IGFBP5-mediated suppression of the IGF pathway, although final birthweights are still low in pre-eclamptic pregnancy.

Mouse model for allogeneic immune reaction against fetus recapitulates human pre-eclampsia.

AIM: We have previously demonstrated that mRNA expression and enzyme activity levels of placental indoleamine 2,3-dioxygenase (IDO), which degrades L-tryptophan and blocks the proliferation of T cells, are significantly low in patients with severe pre-eclampsia. From this observation, we hypothesized that induction of maternal allogeneic immune reaction by reduced IDO activity is one of the causes of pre-eclampsia. METHODS: To examine this hypothesis, we administered an IDO inhibitor to pregnant female mice carrying allogeneic concepti. Since administration of an IDO inhibitor to pregnant mice starting at E4.5 is already reported to cause allogeneic fetal rejection, we modified the regimen and started the administration at E6.5 when the fetus and placenta have already been established. RESULTS: Pregnant mice treated with an IDO inhibitor developed high blood pressure and proteinuria in addition to local circulation impairment in the placenta, which is analogous to the lesions that are characteristic of human pre-eclampsia. In contrast, pregnant mice carrying syngeneic concepti did not manifest such symptoms. CONCLUSIONS: Our findings reveal a pivotal role for IDO activity in the etiology of pre-eclampsia. These data also lend support to the current hypothesis that pre-eclampsia is one of the possible manifestations of a maternal immunological reaction against an allogeneic fetus.

The etiological role of allogeneic fetal rejection in pre-eclampsia.

PROBLEM: It has been demonstrated that allogeneic fetal rejection in normal pregnancy is prevented by placental indoleamine 2,3-dioxygenase (IDO). Further, an immunological etiology has been implicated in pre-eclampsia. METHOD OF STUDY: We examined the differences in placental IDO activity between normal and pre-eclamptic pregnancies. RESULTS: IDO mRNA expression and enzyme activity levels in the placenta were low in patients with severe pre-eclampsia. The enzyme activity also inversely correlates with the blood pressure of the patients. In the placentas from severe pre-eclampsia, IDO immunoreactivity was low, whereas regional T-cell infiltration was observed reciprocally proportional to the IDO activity. CONCLUSION: Our findings implicate a potential role for IDO activity and a maternal immunological reaction against an allogeneic fetus in the etiology of pre-eclampsia.

Establishment and characterization of a human uterine endometrial undifferentiated carcinoma cell line, TMG-L.

A new cell line of human uterine endometrial undifferentiated carcinoma, designated as TMG-L, was established from the metastatic lymph node of 56-year-old patient TMG-L cells have been cultured with Ham's F-12 medium supplemented with 10% FCS and grew as a loosely adherent monolayer with polygonal or spindle-shaped cells exhibiting poor cell-cell contact and piled up against each other, showing a tendency to grow as floating cells. The doubling time of this cell line was about 48 hours, and chromosomal analysis revealed aneuploidy at passage 25. The cells formed tumors in SCID mouse, the histology of which was similar to that of undifferentiated carcinoma component of primary tumor. TMG-L cells showed the loss of expression and membranous localization of either E-cadherin or alpha-catenin, implied corresponding loss of their adhesive function. And this dysfunction implicated the biological aggressive behavior of uterine endometrial undifferentiated carcinoma. This cell line appears to provide a useful system for studying uterine undifferentiated carcinoma in vivo and in vitro.